Regulation of metastasis suppressor NME1 by a key metabolic cofactor coenzyme A
Bess Yi Kun Yu,
Maria-Armineh Tossounian,
Stefan Denchev Hristov,
Ryan Lawrence,
Pallavi Arora,
Yugo Tsuchiya,
Sew Yeu Peak-Chew,
Valeriy Filonenko,
Sally Oxenford,
Richard Angell,
Jerome Gouge,
Mark Skehel,
Ivan Gout
Affiliations
Bess Yi Kun Yu
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Maria-Armineh Tossounian
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Stefan Denchev Hristov
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Ryan Lawrence
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Pallavi Arora
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Yugo Tsuchiya
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom
Sew Yeu Peak-Chew
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, CB2 0QH, United Kingdom
Valeriy Filonenko
Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv, 143, Ukraine
Sally Oxenford
School of Pharmacy, University College London, London, WC1N 1AX, United Kingdom
Richard Angell
School of Pharmacy, University College London, London, WC1N 1AX, United Kingdom
Jerome Gouge
Institute of Structural and Molecular Biology, Birkbeck College, London, WC1E 7HX, United Kingdom
Mark Skehel
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, CB2 0QH, United Kingdom
Ivan Gout
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom; Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv, 143, Ukraine; Corresponding author. Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, United Kingdom.
The metastasis suppressor protein NME1 is an evolutionarily conserved and multifunctional enzyme that plays an important role in suppressing the invasion and metastasis of tumour cells. The nucleoside diphosphate kinase (NDPK) activity of NME1 is well recognized in balancing the intracellular pools of nucleotide diphosphates and triphosphates to regulate cytoskeletal rearrangement and cell motility, endocytosis, intracellular trafficking, and metastasis. In addition, NME1 was found to function as a protein-histidine kinase, 3′-5′ exonuclease and geranyl/farnesyl pyrophosphate kinase. These diverse cellular functions are regulated at the level of expression, post-translational modifications, and regulatory interactions. The NDPK activity of NME1 has been shown to be inhibited in vitro and in vivo under oxidative stress, and the inhibitory effect mediated via redox-sensitive cysteine residues. In this study, affinity purification followed by mass spectrometric analysis revealed NME1 to be a major coenzyme A (CoA) binding protein in cultured cells and rat tissues. NME1 is also found covalently modified by CoA (CoAlation) at Cys109 in the CoAlome analysis of HEK293/Pank1β cells treated with the disulfide-stress inducer, diamide. Further analysis showed that recombinant NME1 is efficiently CoAlated in vitro and in cellular response to oxidising agents and metabolic stress. In vitro CoAlation of recombinant wild type NME1, but not the C109A mutant, results in the inhibition of its NDPK activity. Moreover, CoA also functions as a competitive inhibitor of the NME1 NDPK activity by binding non-covalently to the nucleotide binding site. Taken together, our data reveal metastasis suppressor protein NME1 as a novel binding partner of the key metabolic regulator CoA, which inhibits its nucleoside diphosphate kinase activity via non-covalent and covalent interactions.
